ArticleFEMS microbiology letters2019
CRISPR-Cas9-mediated genome editing in vancomycin-resistant Enterococcus faecium.
Article in FEMS microbiology letters, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 35 citations in OpenAlex.
- Identification of essential genes for conjugative transfer in antimicrobial resistance-associated pELF-type linear plasmids of opportunistic pathogen Enterococcus faecium.PLoS pathogens · 2026Article
- Review
- Cell Death Mechanisms inPathogens (Basel, Switzerland) · 2025Review
- CRISPR-Cas systems in enterococci.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2024Review
- Transcription modulation of pathogenic streptococcal and enterococcal species using CRISPRi technology.PLoS pathogens · 2024Review
- Colonization of vancomycin-resistantFEMS microbes · 2024Article
- Recent advances in gene-editing approaches for tackling antibiotic resistance threats: a review.Frontiers in cellular and infection microbiology · 2024Review
- CRISPR-Cas Systems in the Fight Against Antimicrobial Resistance: Current Status, Potentials, and Future Directions.Infection and drug resistance · 2024Review
- Recent Advances in Understanding the Molecular Mechanisms of Multidrug Resistance and Novel Approaches of CRISPR/Cas9-Based Genome-Editing to Combat This Health Emergency.International journal of nanomedicine · 2024Review
- Application of CRISPR-Cas System to Mitigate Superbug Infections.Microorganisms · 2023Review
- CRISPR-Cas-Based Antimicrobials: Design, Challenges, and Bacterial Mechanisms of Resistance.ACS infectious diseases · 2023Review
- CRISPR-Based Gene Editing inPharmaceuticals (Basel, Switzerland) · 2023Review
- Application of CRISPR-Cas system in the diagnosis and therapy of ESKAPE infections.Frontiers in cellular and infection microbiology · 2023Review
- Barriers to genetic manipulation of Enterococci: Current Approaches and Future Directions.FEMS microbiology reviews · 2022Review
- Characterization of CRISPR-Cas systems inMicrobial genomics · 2022Article
- Recent progress on n-butanol production by lactic acid bacteria.World journal of microbiology & biotechnology · 2021Review
- Let Me Upgrade You: Impact of Mobile Genetic Elements on Enterococcal Adaptation and Evolution.Journal of bacteriology · 2021Review
- RecT Recombinase Expression Enables Efficient Gene Editing inApplied and environmental microbiology · 2021Article
- CRISPR-Cas, a Revolution in the Treatment and Study of ESKAPE Infections: Pre-Clinical Studies.Antibiotics (Basel, Switzerland) · 2021Review
- Harnessing CRISPR-Cas9 for Genome Editing in Streptococcus pneumoniae D39V.Applied and environmental microbiology · 2021Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Gram-positive bacterium Enterococcus faecium is becoming increasingly prevalent as a cause of hospital-acquired, antibiotic-resistant infections. A fundamental part of research into E. faecium biology relies on the ability to generate targeted mutants but this process is currently labour-intensive and time-consuming, taking 4 to 5 weeks per mutant. In this report, we describe a method relying on the high recombination rates of E. faecium and the application of the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas9 genome editing tool to more efficiently generate targeted mutants in the E. faecium chromosome. Using this tool and the multi-drug resistant clinical E. faecium strain E745, we generated a deletion mutant in the lacL gene, which encodes the large subunit of the E. faeciumβ-galactosidase. Blue/white screening using 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal) could be used to distinguish between the wild-type and lacL deletion mutant. We also inserted two copies of gfp into the intrinsic E. faecium macrolide resistance gene msrC to generate stable green fluorescent cells. We conclude that CRISPR-Cas9 can be used to generate targeted genome modifications in E. faecium in 3 weeks, with limited hands-on time. This method can potentially be implemented in other Gram-positive bacteria with high intrinsic recombination rates.
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Registered trials
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